PJB-2020-1516
Gene cloning, recombinant expression and purification of a rice germin like protein (OsRGLP1)
Syed Irtiza Hussain Shah Gardezi, Dure Shahwar, Zaigham Abbas and Syed Muhammad Saqlan Naqvi
Abstract
Germin-like proteins (GLPs) are plant glycoproteins, which are included in cupin superfamily. These proteins show high similarity in sequence but diversity in function and are reportedly involved in conferring biotic and/or abiotic stress tolerance to plants. In order to fully characterize the protein and establish structure-function relationship, it is important to obtain purified protein. Here we report the molecular cloning and recombinant expression of a rice germin like protein 1 (OsRGLP1) in E. coli. Expression vectors pET20b and pET14b carrying C-terminus His tag were used for cloning, optimal expression, and subsequent purification. OsRGLP1 expression was confirmed through western blotting using anti-His monoclonal primary antibody. OsRGLP1 was further purified using Immobilized metal affinity chromatography (IMAC) and size exclusion chromatography. The recombinant protein will be useful for characterizing the protein and raising antibodies for further applications.
To Cite this article:
Gardezi, S.I.H.S. D. Shahwar, Z. Abbas and S.M.S. Naqvi. 2020. Gene cloning, recombinant expression and purification of a rice germin like protein (OsRGLP1). Pak. J. Bot., 52(2): DOI: http://dx.doi.org/10.30848/PJB2020-2(41)
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